Wenjing Decoction, a traditional Chinese medicine known for its blood-regulating effects, is primarily used to treat gynecological disorders in women. Despite its widespread clinical use, there remain gaps in the quality control of Wenjing Decoction, particularly in the comprehensive characterization of its volatile components, which are crucial to its therapeutic efficacy but often overlooked in conventional analyses. To address this issue and provide a more thorough chemical profile of Wenjing Decoction, we applied two complementary analytical methods: offline two-dimensional liquid chromatography coupled with high-resolution mass spectrometry (LCHRMS) and gas chromatography-mass spectrometry (GC-MS). Using LC-HRMS, we successfully characterized 401 non-volatile compounds, which were classified into terpenoids, flavonoids, organic acids, phenylpropanoids, and lactones. GC-MS analysis identified 129 volatile components, including sesquiterpenoids, monoterpenoids, alcohols, and esters. These findings fill a critical gap in the systematic characterization of volatile components in Wenjing Decoction and provide a comprehensive chemical profiling approach for complex traditional Chinese medicine formulations. The results not only support the identification of potential quality control markers but also contribute to the standardization and quality assurance of Wenjing Decoction in clinical practice.

Gastrodia elata Blume is a vital herb requiring precise subtype discrimination for market authentication and traceability. However, the current analytical focus on small molecules limits the extraction of highly discriminative polysaccharide features, making chemometric authentication under class-imbalanced conditions highly challenging. To address these limitations, we established a robust multi-detector size-exclusion chromatography platform utilizing a 5 mM NH₄Ac mobile phase to induce a mixed-mode separation, effectively resolving the cryptic structural heterogeneities and multidimensional properties of the bioactive polysaccharides. Furthermore, we systematically evaluated nine machine learning algorithms, comparing hierarchical versus direct architectures to mitigate class imbalance. For multimodal data fusion, we assessed chromatographic profile, physicochemical expert, and fused feature strategies, utilizing PCA, RFECV, and SFM for dimensionality reduction. Results demonstrated that the hierarchical architecture successfully resolved class imbalance. The KNN model utilizing PCA-reduced fused features achieved optimal performance (Accuracy: 0.8927, F1-Macro: 0.8515). More importantly, interpretable analysis using SHapley Additive exPlanations and PCA loadings revealed a robust chemical decision mechanism: the model prioritizes macroscopic molecular weight (PC1) to identify Gastrodia elata Bl. f. glauca S. Chow (WTM), while utilizing microscopic structural details (PC3/PC5) to distinguish Gastrodia elata Bl. f. elata (HTM) and their hybrids (WHTM). This interpretable chemometric strategy enables precise Gastrodia elata subtype authentication and offers a reliable reference for analyzing polysaccharide-rich herbs.

Areca nut (Areca catechu L.) is a widely consumed food and medicinal plant with both nutritional value and documented health risks. To comprehensively characterize its chemical composition and spatial distribution, we developed an integrated analytical platform combining solid-phase extraction (SPE), offline two-dimensional liquid chromatography with high-resolution mass spectrometry (2D-LC-HRMS), and desorption electrospray ionization mass spectrometry imaging (DESI-MSI). This approach enriched trace constituents while minimizing matrix interference, enabling the identification of 323 compounds-including terpenes and triterpenoids (89), alkaloids (52), flavonoids (31), lignans (17), and steroids (12). A pH-orthogonal separation system (alkaline and acidic modes) significantly improved resolution of structurally similar, low-abundance compounds. Spatial analysis via DESI-MSI revealed three distribution patterns: seed-specific enrichment, pericarp enrichment, and uniform distribution. By integrating compositional and spatial data, this study offers new insights into the pharmacological basis, potential toxicity mechanisms, and safe applications of areca nut in food systems.

Gastrodia elata Blume (GE), a highly regarded traditional Chinese medicinal herb, is widely recognized for its diverse pharmacological activities. Current quality control primarily relies on small molecule biomarkers, yet critical research gaps persist in understanding the complex interactions of the key ingredient, Gastrodia elata polysaccharides (GEP), with bioactivity-related quality due to analytical challenges. To overcome the limitations of traditional non-specific degradation methods, this study established a novel multidimensional quality control strategy using optimized α-amylase-assisted Fenton's initiation toward defined oligosaccharide groups (FITDOG) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous qualitative and quantitative characterization of specific GEP domains. Analysis of 205 batches across varying grades and origins identified α-1,4/1,6-glucans (0.3-40%) as robust, grade-dependent markers. Integration of these macromolecular profiles with small-molecule markers into an Extreme Gradient Boosting (XGBoost) model, superior cultivar discrimination was achieved (Area Under the Curve, AUC > 0.89), while SHapley Additive exPlanations (SHAP) analysis highlighting the predictive significance of the glucan content. Analytical findings were biologically validated in lipopolysaccharide (LPS)-stimulated human microglial clone 3 cells (HMC3), demonstrating that elevated levels of these α-1,4/1,6-glucans significantly correlated with enhanced cell viability (p < 0.05) and anti-neuroinflammatory regulation, characterized by suppression of IL6 and upregulation of TGFB1. This study proposes α-1,4/1,6-glucans as novel quality markers for GE and establishes a robust analytical framework that integrates chemical profiling and biological validation. This advancement enhances the precision of quality control in traditional Chinese medicine and underscores the potential of polysaccharide-driven strategies for the standardization of medicinal herbs.

A novel and efficient one-pot synthesis of /3-carboline derivatives through a tandem Pictet-Spengler/oxidative aromatization sequence using calcium hypochlorite (Ca(ClO)2) as a mild oxidant has been reported. This methodology enables the direct conversion of readily available tryptamine derivatives and aldehydes in methanol to afford the corresponding /3-carboline products in moderate to excellent yields (34-92 %). Notably, this protocol eliminates the need for isolation of the intermediate 1,2,3,4-tetrahydro-/3-carbolines, thereby significantly streamlining the synthetic process. The operational simplicity, good functional group tolerance, and avoidance of harsh reaction conditions make this method a practical approach for the synthesis of /3-carboline scaffolds, which are privileged structures in medicinal chemistry.

A green and efficient method for synthesizing imidazolidin-4-one derivatives catalyzed by thiamin hydrochloride (VB1) has been developed. This protocol involves the reaction of alpha-amino amides with carbonyl compounds (aldehydes or ketones) in the presence of VB1 (5 mol%) in EtOH, affording the target products in 50-85% yields. The reaction demonstrates broad substrate compatibility, accommodating aromatic aldehydes, heteroaromatic aldehydes, alkyl aldehydes, and alkyl ketones. Notably, estrone as a ketone substrate successfully undergoes condensation, yielding the corresponding product in 72% yield. The VB1 catalyst offers advantages such as short reaction times, high efficiency, excellent environmental friendliness, and recyclability, maintaining performance over at least three cycles. [GRAPHICS]

BACKGROUND:Keratinocytes form the skin's first line of defense, not only serving as a physical barrier but also actively communicating with immune cells and sensory neurons. OBJECTIVE:This study elucidated the molecular mechanisms by which keratinocytes contribute to barrier dysfunction and neuroimmune activation in atopic dermatitis (AD). METHODS:Cannabinoid receptor 2 (CB2R) expression was assessed by RNA sequencing, quantitative reverse transcription PCR (qRT-PCR), RNAscope fluorescence, and Western blot analysis. Pharmacologic activation/inhibition or keratinocyte-specific deletion of CB2R was used to determine its role in AD-associated itch and inflammation. Behavioral assays, immunofluorescence, and qRT-PCR were used to identify downstream signaling components. RESULTS:CB2R expression was upregulated in the epidermis of MC903-induced AD. CB2R activation alleviated scratching and skin pathology, whereas keratinocyte-specific CB2R deletion exacerbated both. CB2R suppressed adenylate cyclase 3 activity, reducing cAMP levels and downstream Epac1 activation, thereby limiting IL-33 production. This CB2R-cAMP-Epac1-IL-33 axis regulated epidermal hyperplasia, dermal neutrophil infiltration, and chronic itch. CONCLUSION:The keratinocyte CB2R-cAMP-Epac1 axis integrates epithelial, neural, and immune signaling to drive AD pathology, representing a potential therapeutic target for chronic itch and skin inflammation.

Gastrodia elata Blume (G. elata) has been utilized as homologous origin in medicine and food for over 2000 years. Gastrodia elata polysaccharides (GEP), the key bioactive components, have attracted increasing attention owing to their natural origin, potent bioactivities and low toxicity. Despite significant progress, multiple critical issues still need to be addressed. Current studies often summarize structural characterization and activities separately, with limited focus on advanced structures and structure-activity relationships. Moreover, the insufficient emphasis of GEP in the quality control of G. elata in the current pharmacopoeia has led to persistent challenges, including batch variability and lack of standardization of GEP. This review systematically summarizes recent advancements in the extraction, purification, and structural characterization of GEP, specifically to summarize the advanced structure of GEP and to introduce new techniques for structural elucidation. While we highlight relationships between structures and activities, and discuss their potential role in improving G. elata quality control. Looking forward, a dual chemical-biological quality control framework is proposed, alongside multidisciplinary approaches such as metabolomics, synthetic biology, and artificial intelligence (AI) to promote standardized production and clinical translation of GEP, thereby facilitating their broader application in medicine and functional foods.

The rapid screening of bioactive constituents within traditional Chinese medicine (TCM) presents a significant challenge to researchers. Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents. To address this limitation, a fishing strategy based on offline two-dimensional liquid chromatography (2D-LC) combined with surface plasmon resonance (SPR) was utilized to screen bioactive trace components targeting peroxiredoxin 3 (PRDX3), using Uncaria alkaloids (UAs) as a case study. Initially, an orthogonal preparative offline 2D-LC system combining a positively charged C18 column and a conventional C18 column under disparate mobile phase conditions was constructed. To fully reveal the trace alkaloids, 13 2D fractions of UAs were prepared, and their components were characterized using mass spectrometry (MS). Subsequently, employing PRDX3 as the targeting protein, a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether (GSM) serving as a positive control for binding. Employing this refined strategy, 29 candidate binding alkaloids were fished from the 13 2D fractions. Notably, combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone. Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids, with isovallesiachotamine (IV), corynoxeine N-oxide (CO-N), and cadambine (CAD) demonstrating the highest affinity for PRDX3. Their interactions were further validated through molecular docking analysis. Subsequent intracellular H2O2 measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H2O2 clearance. In conclusion, this study introduced an innovative strategy for the identification of active trace components in TCM. This approach holds promise for accelerating research on medicinal components within this field.

Gastrodiae Rhizoma (Tianma, GR), the dried tuber of Gastrodia elata Bl., is a valuable herb for medicinal and dietary applications. Current markets feature three key cultivars: G. elata f. glauca (WTM), G. elata f. elata (HTM), and their hybrid (WHTM), with significant price variations yet lacking defined chemical authentication markers. Here, we implemented a pseudotargeted LC-MS/MS metabolomics strategy employing 534 optimized MRM transitions. This strategy was coupled with multivariate analysis - unsupervised principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) - of 40 authenticated batches, to decipher cultivar-specific chemical fingerprints. The approach identified 88 statistically validated markers, with 14 pivotal discriminators-primarily 4-Hydroxybenzyl alcohol derivatives-exhibiting distinct cultivar distribution patterns. This chemical blueprint serves as a tool for authentication in quality control and supports regulatory standard establishment for GR products.

A novel and efficient one-pot synthesis of β-carboline derivatives through a tandem Pictet-Spengler/oxidative aromatization sequence using calcium hypochlorite (Ca(ClO)2) as a mild oxidant has been reported. This methodology enables the direct conversion of readily available tryptamine derivatives and aldehydes in methanol to afford the corresponding β-carboline products in moderate to excellent yields (34–92 %). Notably, this protocol eliminates the need for isolation of the intermediate 1,2,3,4-tetrahydro-β-carbolines, thereby significantly streamlining the synthetic process. The operational simplicity, good functional group tolerance, and avoidance of harsh reaction conditions make this method a practical approach for the synthesis of β-carboline scaffolds, which are privileged structures in medicinal chemistry.

Andrographis paniculata (Burm. f.) and its products have a long history of medicinal use in Asia. A. paniculata products are mainly made from the root extraction of stems, leaves and parts, but there may be differences in the proportion of different parts and different harvest times, which ultimately leads to certain differences in product quality. In this study, the chemical components and non-targeted metabolomics were characterized, and the characteristic compounds in different parts of A. paniculata at various growth stages were analyzed. By utilizing polygonal mass defect filtering, precursor ion lists, and a self-built compound library, a total of 225 components were identified in A. paniculata. Notably, spermidine derivatives and phosphatidylcholines were reported for the first time in this plant species. In total, 41 differential components were identified in different parts of A. paniculata. These findings provide scientific evidence for the selection of quality markers in A. paniculata and its products.

The complex pathological mechanisms of non-alcoholic fatty liver disease (NAFLD) are closely related to dysregulated lipid metabolism, and the therapeutic effects of the traditional Chinese medicine Zexie-Baizhu Decoction (AA) on NAFLD have been gaining increasing attention. However, research into altered lipid metabolism, especially fatty acids, in NAFLD and the intervention of AA faces technical challenges, especially in the precise quantitative analysis of fatty acids in biological samples. The high complexity of biological matrices, particularly after drug intervention, greatly increases the difficulty of detection. Therefore, this study innovatively developed a simple and economical stable isotope derivatization technique by synthesizing d6N,N-dimethylethylenediamine (d6-DMED) in the laboratory, establishing a simple and economical method for fatty acid quantification. This method employs a chemical reaction under low-temperature conditions to ensure the efficient synthesis of d6-DMED. Using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry technique (UHPLC-MS/MS), combined with optimized chromatographic separation conditions and dynamic multiple reaction monitoring mode, the study established a highly sensitive detection method for 35 fatty acid derivatives. Methodological evaluation showed that the limits of quantification ranged from 0.002 to 0.060 μM, with high linearity of R² > 0.995. Additionally, the relative recovery rates were between 93.14% and 106.63%. To further demonstrate the feasibility of this method for fatty acid quantification, it was applied to measure fatty acids in multiple tissues in a mouse NAFLD model, as well as the effects of AA intervention on fatty acid metabolism. This rapid, simple, and cost-effective detection method not only enhances the understanding of NAFLD mechanisms but also provides a new strategy for evaluating the biological complex system after drug intervention.

During the coronavirus reproduction process, 3-chymotrypsin-like protease (3CLpro) and papain-like protease (PLpro) are accountable for the fragmentation of two polyprotein precursors (pp1a/pp1ab) into substructural proteins. These two proteins are vital for the replication and transcription of the viral genome. Therefore, 3CLpro is a key protein and target for the design of coronavirus inhibitors. In previous studies, we found that betulinic acid has an inhibitory effect on 3CLpro, with 51.5 % inhibition of 3CLpro at 20 mu M. Then, series of betulinic acid derivatives were designed, synthesized, and evaluated for their inhibition activities. The results showed that BA02 and BA05 showed significant inhibitory activity on 3CLpro with inhibitory rates of 78.1 % and 82.5 % at 20 mu M, respectively. Further evaluation of these two compounds shows that their IC50 values are 7.22 +/- 0.14 mu M and 6.40 +/- 0.14 mu M, respectively.

BACKGROUND:Energy deficiency is the characteristic of chemotherapy-induced cachexia (CIC) which is manifested by muscle wasting. glycolysis, tricarboxylic acid (TCA) cycle, and lipid metabolism are central to muscle bioenergy production, which is vulnerable to chemotherapy during cancer treatment. Recent investigations have spotlighted the potential of Shenqi Fuzheng injection (SQ), a Chinese proprietary medicine comprising Radix Codonopsis and Radix Astragali, in alleviating CIC. However, the specific effects of SQ on muscle energy metabolism remains less explored.PURPOSE AND METHODS:Here, we integrated transcriptomics, spatial metabolomics, gas chromatography-mass spectrometry targeted quantitative analysis, and transmission electron microscopy techniques, combined with Seahorse live-cell metabolic analysis to reveal the changes in genes and pathways related to energy metabolism in the CIC model and SQ's protective effects at molecular and functional levels.RESULTS:Our data showed that chemotherapeutic agents caused glycolysis imbalance, which further leads to metabolic derangements of TCA cycle intermediates. SQ maintained glycolysis balance by facilitating pyruvate fluxing to mitochondria for more efficient bioenergy production, which involved a dual effect on promoting functions of mitochondrial pyruvate dehydrogenase complexes and inhibiting lactate dehydrogenase for lactate production. As a result of the sustained pyruvate level achieved by SQ administration, glycolysis balance was maintained, which further led to the preservation of mitochondrial integrity and function of electron transport chain, thereby, ensuring the normal operation of the TCA cycle and the proper synthesis of adenosine triphosphate (ATP). The above results were further validated using the Seahorse live-cell assay.CONCLUSION:In conclusion, our study highlights SQ as a promising strategy for CIC management, emphasizing its ability to harmonize the homeostasis of the muscle bioenergetic profile. Beyond its therapeutic implications, this study also offers a novel perspective for the development of innovative treatments in the realm of herbal medicine.

INTRODUCTION:Amomum fruit, also known as Sharen, serves as both a functional food and a traditional Chinese medicine with significant pharmacological activities. However, there are three botanical origins of Amomum fruit: Amomum villosum Lour. (AVL), Amomum villosum Lour. var. xanthioides T. L. (AVX), and Amomum longiligulare T. L. Wu (ALW). OBJECTIVE:Conducting a comprehensive chemical composition analysis of Amomum fruit from three botanical origins aims to identify potential differences in metabolic characteristics. METHODS:To annotate the metabolic characteristic ions of multi-origin Amomum fruit, we employed metabolomic techniques, including ultra-high-performance liquid chromatography (LC) coupled with linear ion trap-Orbitrap-tandem mass spectrometry (MS) and gas chromatography-MS, in conjunction with feature-based molecular networking technology. Additionally, chemometrics was utilized to examine the correlations between the various botanical origins. RESULTS:A total of 201 non-volatile and 151 volatile metabolites were annotated, and most of the proanthocyanidins and flavonoids were identified by feature-based molecular networking. Additionally, 61 non-volatile and 45 volatile feature ions were screened out for classification. Principal component analysis, orthogonal projection to latent structures discrimination analysis, and heat map analysis were employed to clearly distinguish the metabolite profiles of Amomum fruit from different origins. Hierarchical clustering analysis indicated that proanthocyanidins C1 and C2, as well as proanthocyanins oligomers, show significant differential expression between AVX and AVL, which could be the new quality markers for the classification. CONCLUSION:Classification of the botanical origin of Amomum fruit based on LC-MS characteristic ions proved to be more advantageous. This study introduces new strategies and technical support for the quality control of Amomum fruit and facilitates the identification of unknown compounds for future research.

ETHNOPHARMACOLOGICAL RELEVANCE:Zexie-Baizhu Decoction (AA), a Chinese Classical Formula composed of Alisma orientalis (Sam.) Juzep. and Aractylodes Macrocephala Koidz in the specific ratio of 5:2, has a long history of use in treating metabolic disorders. Recent studies have demonstrated AA's ameliorative effects on non-alcoholic fatty liver disease (NAFLD); however, the mechanism underlying its action on the gut and adipose tissue, key regulators of metabolism, have not been fully explored. AIM OF THE STUDY:This study aimed to investigate the mechanisms by which AA regulates the homeostasis of gut and adipose tissue in NAFLD. MATERIALS AND METHODS:AA (1500 mg/kg/day) or vehicle was administrated to the high-fat diet-induced and normal chow-fed mice (C57BL/6J). Plasma, the liver, gut microbiota, bile acids, and short-chain fatty acids in the gut, were systematically investigated. RNA sequencing analysis, reverse transcription quantitative real-time PCR, and Western Blotting were performed on the epididymal white adipose tissues (eWAT) to explore AA's influence on NAFLD. Lipidomics of the liver and eWAT were analyzed by liquid chromatography-mass spectrometry and desorption electrospray ionization mass spectrometry imaging. RESULTS:Our study demonstrated that AA administration effectively alleviated liver injury induced by NAFLD, as evidenced by reduced hepatic fat accumulation and inflammation. Mechanistically, AA modulated the composition of the gut microbiota, promoting the growth of beneficial bacteria such as Akkermansia muciniphila and restoring the balance between Firmicutes and Bacteroidetes. Furthermore, AA regulated the levels of bile acids and short-chain fatty acids in the intestine, plasma, and liver. Correspondingly in the eWAT, AA administration activated bile acid receptor (Gpbar1) and short-chain fatty acid receptor (Ffar2), facilitating lipid breakdown and attenuating triglyceride accumulation. Transcriptome analysis revealed that AA influenced gene expression related to fatty acid metabolism, thermogenesis, insulin resistance, AMPK signaling, and the tricarboxylic acid (TCA) cycle, thereby improving NAFLD at the transcriptional level. Additionally, AA treatment significantly altered the lipid composition in the liver, reducing levels of diacylglycerols, triacylglycerols, phosphatidylserines, and cholesterol esters, while increasing levels of phosphatidic acids, phosphatidylethanolamines, and sphingomyelins. CONCLUSION:Our study builds a connection between the gut and adipose tissue to understand the mechanism of AA on alleviating NAFLD, providing new insights into the development of targeted therapies for this condition.

Angelicae sinensis radix is commonly used in Chinese medicine and is also widely used as a spice and condiment in many countries. Due to the different geographical sources and processing methods, the contents of chemical components are different and affect the quality of Angelica sinensis radix. A simple and efficient method was established to comprehensively evaluate the quality of Angelica sinensis radix by combining HPLC fingerprint and single-standard multi-component quantitative analysis. With ferulic acid as a single marker, the contents of eight main active ingredients, including ferulic acid, senkyunolide I, senkyunolide H, coniferyl ferulate, E -ligustilide, E -butylidenephthalide, Z -ligustilide, and Z- butylidenephthalide, were simultaneously determined in 30 min by reliable relative correction factor. The overall chemical profiles of Angelica sinensis radix were compared by fingerprint similarity evaluation, the specific content level of each component was compared by simultaneous content determination of multiple components of a single standard, and the specific different components of Angelica sinensis radix before and after processing were analyzed by chemometrics. The established method was successfully applied to the quality evaluation of 44 batches of Angelica sinensis radix from 4 different origins and 45 batches of Angelica sinensis radix with different processing methods. This study proposed a simple, rapid, low-cost, and reliable quality comprehensive evaluation method for ASR from different origins and processing methods, which may be readily transferred and applied to other industries.

INTRODUCTION:Lipid metabolism participates in various biological processes such as proliferation, apoptosis, migration, invasion, and maintenance of membrane homeostasis of prostate tumor cells. Bufadienolides, the active ingredients of Chansu, show a robust anti-proliferative effect against prostate cancer cells in vitro, but whether bufadienolides could regulate the lipid metabolism in prostate cancer has not been evaluated.OBJECTIVES:Our study explored the regulatory effects of bufadienolides on lipid metabolism in human prostate carcinoma cells (PC-3).METHODS:Untargeted lipidomics and transcriptomics were combined to study the effect of different bufadienolides interventions on lipid and gene changes of PC-3 cells. The key genes related to lipid metabolism and prostate cancer development were verified by qPCR and western blotting.RESULTS:Lipidomic analysis showed that the active bufadienolides significantly downregulated the content of long-chain lipids of PC-3 cells. Based on transcriptomic and qPCR analyses, many genes related to lipid metabolism were significantly regulated by active bufadienolides, such as ELOVL6, CYP2E1, GAL3ST1, CERS1, PLA2G10, PLD1, SPTLC3, and GPX2. Bioinformatics analysis of the Cancer Genome Atlas database and literature retrieval showed that elongation of very long-chain fatty acids protein 6 (ELOVL6) and phospholipase D1 (PLD1) might be important regulatory genes. Western blot analysis revealed that active bufadienolides could downregulate PLD1 protein levels which might promote anti-prostate cancer effect.CONCLUSIONS:All these findings support that bufadienolides might induce lipid metabolic remodeling by regulating long-chain lipids synthesis and phospholipid hydrolysis to achieve an anti-prostate cancer effect, and PLD1 would probably be the key protein.